Thyrotropin-receptor-mediated diseases: a paradigm for receptor autoimmunity

Autoantibodies to the thyrotropin receptor (TSHR) can act as thyrotropin agonists or antagonists, or can cause thyroid hypertrophy. Neither the autoantibody-binding sites on the TSHR nor the intracellular mechanisms by which the autoantibodies mediate their diverse functional effects are completely understood. This article reviews how cloning of the TSHR has contributed to our understanding of its structure and function, and has allowed induction of experimental autoimmunity to the TSHR.     

Impact of Zinc oxide nanoparticles on eggplant (S. melongena): studies on growth and the accumulation of nanoparticles

The increasing use of nanoparticles and their occurrence in the environment has made it imperative to elucidate their impact on the environment. Although several studies have advanced the authors’ understanding of nanoparticle–plant interactions, their knowledge of the exposure of plants to nanoparticles and their effects on edible crop plants remain meager and is often paradoxical. The aim of this study was to increase their knowledge on the effect of zinc oxide (ZnO) nanoparticles on eggplant seed germination and seedling growth. ZnO nanoparticles had a negative effect on the growth of eggplant in plant tissue‐culture conditions, as the growth of seedlings decreased with the increase in the concentration of ZnO nanoparticles. In contrast, ZnO nanoparticles enhanced eggplant growth under greenhouse conditions. The accumulation of ZnO nanoparticles in various parts of eggplant was observed through scanning electron microscopy of both plant tissue‐culture and greenhouse‐raised eggplant seedlings. To the best of their knowledge, this is the first study to report on ZnO nanoparticle accumulation in eggplant and its effect on seed germination and seedling growth.Inspec keywords: crops, zinc compounds, scanning electron microscopy, II‐VI semiconductors, nanoparticles, agriculture, cellular biophysics, nanofabricationOther keywords: plant tissue‐culture, greenhouse‐raised eggplant seedlings, ZnO nanoparticle accumulation, seedling growth, ZnO nanoparticles, nanoparticle–plant interactions, zinc oxide nanoparticles, eggplant seed germination, eggplant growth, ZnO     

Influence of adjuvants on the induction of autoantibodies to the thyrotropin receptor

To determine the influence of adjuvant on the induction of antibodies to thyrotropin receptor (TSHR), we immunized BALB/c mice with a extracellular domain of the TSHR (ETSHR) protein in complete Freund's adjuvant (CFA), Titer Max (TM) and Gerbu. Similarly, control groups of mice were immunized with bovine serum albumin (BSA) in each of the different adjuvants. As determined by ELISA, ETSHR given along with CFA elicited high titers of antibodies to ETSHR which were mainly restricted to the IgG1 subclass. Mice immunized with ETSHR in TM also developed high titers of anti-ETSHR antibodies but had higher levels of both IgG1 and IgG2a. However, immunization with ETSHR in Gerbu resulted in low titers of antibodies, restricted to IgG1 subclass. Immunization of mice with BSA in each of the three adjuvants induced higher antibody titers to BSA. The subclass of antibodies in mice immunized with BSA in CFA and TM were predominantly IgG1 and IgG2a with lower levels of IgG2b, whereas in Gerbu treated group, antibody to BSA was restricted to IgG1 subclass. Analysis of specificity of antibodies against ETSHR, in mice immunized with ETSHR, revealed that irrespective of the adjuvant used, the dominant reactivity was against peptide 1 (AA 22-41) with weaker reactivity against several other. peptides. The only exception was in mice immunized with ETSHR in TM which also showed significant reactivity against peptide 23 (AA 352-371). Mice immunized with the ETSHR in CFA or in TM showed elevated levels of serum TSH binding inhibitory immunoglobulins (TBII). However, mice immunized with ETSHR in Gerbu, which had lower titers of antibodies to ETSHR, showed normal TBII levels. These studies showed that adjuvant composition could influence the titer, subclass and fine specificity of antibodies to ETSHR which in turn could affect the development of TBII activity.     

Influence of thermal pressure and temperature profile approximations in thermohydrodynamic analysis of surface-textured noncircular journal bearing

The present study aims to study the thermohydrodynamic performance of noncircular journal bearings lubricated with multigrade oil. Experimental investigations are carried out on phosphorous bronze hydrodynamic journal bearing with a surface textured at different loading conditions with multigrade oil (MAK 2T oil) as lubricant at different constant rotational speeds (i.e., 1000–4000 rpm) for studying the dynamic characteristics. The journal-bearing's thermodynamic behavior is analyzed by simplifying modified Reynolds and energy equations to obtain surface-textured bearing performance characteristics, such as load-carrying capacity, mass flow rate, and power loss. The noncircular bearings suggest several design parameters, such as a tilt angle for designers. By considering thermal effects for MAK 2T oil lubricant, the requirements of a specific application can be fulfilled.     

Flow cytometric analyses of antibody binding to Chinese hamster ovary cells expressing human thyrotropin receptor

To develop a method that can be used to directly detect binding of antibodies to TSH receptor (TSHr), we employed Chinese hamster ovary (CHO) cells permanently transfected with a human TSHr complementary DNA (CHOR). These cells showed increased cAMP production when treated with either human TSH or thyroid-stimulating antibodies and decreased TSH-mediated cAMP production when treated with stimulation-blocking antibodies. We employed flow cytometry and rabbit antibodies against the extracellular domain of the TSHr (ETSHr) to test whether these cells can be used to directly detect and quantitate the binding of anti-TSHr antibodies. Rabbit anti-ETSHr bound specifically to CHOR cells, and the binding could be blocked with purified ETSHr. To test the feasibility of using these cells for epitope mapping, we tested the binding of rabbit antibodies raised against several synthetic TSHr peptides. Rabbit antipeptide 92 (amino acids 12-30) and 91 (amino acids 32-46) showed little or no binding to the CHOR cells. In contrast, antibodies raised against peptides 93 (amino acids 316-330), 95 (aa 325-345), 3A (aa 357-372), 367 (aa 367-386), and 1B (aa 362-376) showed significant binding to the CHOR cells. The specificity of binding of antipeptide antibodies was demonstrated by a complete inhibition of binding by corresponding peptides. When TSH-binding inhibitory Ig-positive sera from 15 patients with hyperthyroidism were tested, 8 of them showed specific binding to the CHOR cells compared to their relative binding to normal CHO cells; sera from all normal individuals tested did not exhibit specific binding to CHOR cells. These studies showed the usefulness of CHOR cells and flow cytometry in epitope mapping using sera with known specificities and the potential usefulness of the technique to detect anti-TSHr antibodies in patient sera.     

Purification and characterization of a recombinant human thyroid peroxidase expressed in insect cells

Thyroid peroxidase (TPO) is an essential enzyme for thyroid hormone biosynthesis and is an autoantigen against which antibodies are found in a number of autoimmune thyroid disorders. Large quantities of pure TPO are essential for understanding its structure and role in normal thyroid function and thyroid diseases. In this study, we describe the production of human TPO (hTPO) using a baculovirus expression vector in insect cells. TPO was sequentially extracted from insect cells using various buffers and the protein was purified to homogeneity on a C4 reversed-phase semipreparative column using high-performance liquid chromatography. The purified protein was identified as hTPO by enzyme-linked immunosorbent assay, Western blot, and amino acid sequence analyses. Carbohydrate analysis of the recombinant hTPO showed that the protein is glycosylated and mannose is the major oligosaccharide. We have extended the carbohydrate analysis by establishing the occurrence of N-acetyl galactosamine which suggested that the recombinant hTPO might contain O-glycosyl moieties. Purified hTPO reacted specifically with sera from patients with Hashimoto's thyroiditis. Crude as well as purified hTPO did not show any enzymatic activity when produced in Sf9 insect cells grown in serum free medium. In contrast, hTPO produced in the presence of 10% fetal bovine serum containing 1 microgram/ml of haematin was enzymatically active. However, the enzymatic activity of the recombinant hTPO was lower than that often found with hTPO purified from thyroid tissue. Availability of purified hTPO in relatively large quantities should allow further structural and immunological studies.     

Dietary agents in the prevention of alcohol-induced hepatotoxicty: preclinical observations

Long term alcohol consumption is one of the important causes for liver failure and death. To complicate the existing problem there are no dependable hepatoprotective drugs and a large number of patients prefer using complementary and alternative medicines for treating and managing hepatic complications. Almost 25 centuries ago, Hippocrates, the father of medicine, proclaimed "Let food be thy medicine and medicine be thy food." Exploring the association between diet and health continues even today. Preclinical studies carried out in the recent past have shown that the commonly used dietary agents like Allium sativum (garlic), Camellia sinensis (tea), Curcuma longa (turmeric), Emblica officinalis (Indian gooseberry), Ferula asafoetida (asafoetida), Garcinia cambogia (Malabar tamarind), Glycine max (soyabean), Murraya koenigii (curry leaves), Piper betle (beetle leaf), Prunus armeniaca (apricot), Ocimum gratissimum (wild basil), Theobroma cacao (cocoa), Trigonella foenum-graecum (fenugreek) and Vitis vinifera (grapes) protect against ethanol-induced hepatotoxicity. Mechanistic studies have shown that the beneficial effects of these phytochemicals in preventing the ethanol-induced hepatotoxicity are mediated by the antioxidant, free radical scavenging, anti-inflammatory and anti-fibrotic effects. The present review for the first time collates the hepatoprotective effects of these agents and also emphasizes on aspects that need future research to establish their utility in humans.     

Synthesis and rheological characterization of nano-magnetorheological fluid using inverse spinel ferrite (NiFe2O4)

In recent years, the utilization of nanoparticles for nano-magnetorheological fluid (NMRF) synthesis is gaining popularity in automotive applications. From this perspective, the nickel ferrite (NiFe₂O₄) nanoparticles were prepared by gel burning method and characterized using the X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), field emission scanning electron microscopy-energy dispersive X-ray analysis (FESEM-EDX), and vibration sample magnetometer (VSM). The XRD and FTIR results showed the phase formation and characteristic metal–oxygen M–O vibrations. The FESEM images showed quasi-spherical crystallites with considerable agglomeration. The magnetic properties measured showed the ferromagnetic nature of NiFe₂O₄. The nanosized NiFe₂O₄ was used for NMRF preparation and characterization.